CN1662248A - Compositions against inflammatory processes - Google Patents

Abstract

Compositions capable of reducing a risk of inflammation in a mammal(s) are provided. The compositions contain a therapeutically effective amount of a thermally processed (e.g., extruded) plant material that includes alpha-methylene-gamma-butyrolactone (alpha-MGBL) and/or several structurally related parent phytochemicals capable of inhibiting enzymatic and/or transcriptional activity in the mammal, which is believed to treat and/or prevent inflammation. In this regard, the risk of incidence of other disease or ailments which are believed to be caused by inflammation (e.g., chronic inflammation) may be reduced. The phytochemical source can be derived from a single plant material, such as chicory.

Description

Composition against inflammatory processes

The present invention generally relates to compositions. More specifically, the present invention relates to thermally processed (eg extruded) plant material, such as chicory and/or extracts thereof, included to enhance human and animal health. Plant material from, for example, plants of the family Asteracae, which contains the desired amount of a sesquiterpene lactone effective to produce its active product such as α-methylene-γ-butyrolactone (α-MGBL) upon heating , or from other plants containing the same or substantially the same compound, such as coffee or soybeans.

Background of the invention

The need to enhance mammalian health has involved and continues to involve ongoing research efforts and discoveries to prevent and/or treat disease. For example, anger or discomfort may be caused by inflammation in the mammal such as dermatitis, ophthalmia, enteritis, and the like. Additionally, chronic inflammation is generally believed to increase the risk of other diseases or ailments such as osteoarthritis, autoimmune diseases, cancer, and the like. In this regard, it is generally understood that inflammation is triggered by enhanced transcriptional activity of the transcription factor NF-κB, which leads to the expression of proinflammatory and inflammatory enzymes and receptors. More specifically, inflammation can occur due to enhanced enzymatic activity in mammals such as enhanced activity of cyclooxygenases including cyclooxygenase-1, cyclooxygenase-2, and the like.

In general, foods, diets or other sources of nutrition are known to contain a number of ingredients or agents believed to protect humans and animals from disease. For example, oligosaccharides such as inulin and various fructo-oligosaccharides have been reported to have prebiotic effects, such as promoting bifidobacteria and Growth of lactobacillus. See, eg, Gibson et al., Food Microbiology, Vol. 11 (No. 6), pp. 491-498 (1994). Although most of the reported experiments were performed in vitro, it has also been reported that these oligosaccharides have similar effects in human and mouse intestines. In this regard, it is generally believed that the promotion of the growth of bifidobacteria and lactobacilli through the use of oligosaccharides can provide a variety of beneficial effects in animals and humans, such as prevention and/or treatment of dysentery, promotion of growth, enhancement of reproductive capacity, or Other health-enhancing beneficial effects.

Inulin or other dietary ingredients, as discussed above, that are believed to enhance human and animal health, in general, may be derived from plants or other natural sources. For example, inulin is generally known to be purified from plants containing high concentrations of inulin such as chicory, Jerusalem artichoke, leek and asparagus. In this regard, the plant is typically purified or otherwise treated prior to use to enhance the flavor of the plant, such as to eliminate or at least minimize the bitterness typically associated with chicory. See, e.g., U.S. 4,865,852.

Purified plant products are usually prepared by acid or enzymatic hydrolysis. The hydrolyzate is then collected and concentrated to obtain bioactive agents such as inulin. For example, JP 63-309147 discloses ground chicory tubers, partially hydrolyzed with acid and then drying the hydrolyzate with or without neutralization. However, purification of, for example, oligofructosaccharides and inulin can greatly increase the cost of dietary products. Therefore, the use of such dietary products is generally limited to special foods or dietary products for humans and animals.

A need therefore exists for a composition comprising natural ingredients such as heat-processed chicory and/or its extracts which is palatable to humans and animals, which can be produced cheaply and which enhances the health of humans and animals , such as preventing and/or treating inflammation.

Summary of the invention

The present invention relates to compositions useful for enhancing human and animal health, especially for preventing and/or treating inflammation. Compositions of the invention include one or more phytochemicals derived from thermally processed (eg extruded) plant sources such as chicory.

Applicants have demonstrated that chicory root extracts contain one or more phytochemicals that inhibit enzymatic and/or transcriptional activities in mammals, such as those involving cyclooxygenase and those involving NF- κB transcriptional activity. Applicants have also demonstrated that thermally processed (eg extruded) chicory root extracts have enhanced enzyme inhibitory activity involving cyclooxygenase and/or enhanced inhibitory effects involving NF-KB. Applicants demonstrated a relationship between the enhancement of inhibition and the production of the active product, alpha-methylene-gamma-butyrolactone (a-MGBL), during thermal processing.

In this regard, inhibition of such enzymatic/transcriptional activity has been shown to prevent and/or treat inflammation in mammals. By inhibiting the inflammatory process, the risk of developing other diseases or ailments thought to arise from inflammation (eg, chronic inflammation), such as cancer, can be reduced.

To this end, in one embodiment of the present invention there is provided such a nutritional composition. The composition comprises a therapeutically effective amount of plant material comprising one or more heat-processed (eg extruded) phytochemicals capable of inhibiting enzyme activity to prevent and/or treat inflammation in a mammal. Preferably, plant material comprises at least 0.5% by weight.

In one embodiment, the plant material is from Compositae or some other plant such as coffee, soybean, etc. or a combination thereof. Preferably, the plant material is from chicory, lettuce, coffee, soybean or a combination thereof. In one embodiment, the plant material includes chicory extract.

In addition to phytochemicals, the composition may also include other dietary ingredients from plant sources. These dietary ingredients may include any suitable component capable of inhibiting the enzymatic activity associated with cyclooxygenase. In one embodiment, dietary ingredients other than phytochemicals include antioxidants, glucosamine, chondroitin sulfate, omega-3 fatty acids, any suitable other dietary ingredients, and combinations thereof.

The present invention also provides pet food products. The pet food product includes a starch base; and an effective amount of thermally processed (eg, extruded) plant material comprising a phytochemical capable of inhibiting enzyme activity in a mammal thereby reducing the risk of inflammation.

In another embodiment of the present invention, a method of preparing a nutraceutical food product that reduces the risk of inflammation in a mammal is provided. The method comprises the steps of: providing plant material; thermally processing the plant material to form a plant extract comprising one or more phytochemicals capable of inhibiting enzyme activity in a mammal; and processing the plant extract and one or more foods Ingredients to form a nutritious food product comprising at least 1% by weight of plant extracts.

Preferably, the plant extract is processed by defatting of the plant material to form the primary plant extract, and the primary plant extract is subsequently processed by acid hydrolysis with ethyl acetate to form the plant extract.

The present invention also provides the use of a therapeutically effective amount of heat-processed, such as extruded, plant material comprising one or more phytochemicals capable of inhibiting enzyme activity in a mammal, for the preparation of Composition of inflammatory risks. With reduced enzyme activity, the risk of inflammation in the mammal is reduced, thereby reducing the risk of developing other ailments or diseases caused by inflammation, such as osteoarthritis, autoimmune diseases, cancer, and the like.

It is an advantage of the present invention to provide an improved composition useful for reducing the risk of inflammation in mammals. In this regard, the compositions are capable of inhibiting enzymatic activity, the result of which is believed to treat and/or prevent inflammation.

Another advantage of the present invention is to provide an improved composition comprising plant material containing one or more phytochemicals that inhibit, for example, enzymatic activity involving cyclooxygenases, which Thought to prevent and/or treat inflammation in mammals.

Another advantage of the present invention is to provide a method for producing an improved composition comprising plant material that enhances the palatability of the composition while maintaining the presence of plant material that reduces the risk of inflammation in mammals. Enzyme inhibitory properties.

Another advantage of the present invention is to provide a method of treating and/or preventing inflammation in a mammal comprising administering the improved composition.

Other features and advantages of the present invention are described in the following detailed description of the invention, and these other features and advantages will be apparent.

Brief description of the drawings

Figure 1 illustrates the inhibitory effect of COX-2 expression in HT29 cells by α-methylene-γ-butyrolactone (α-MGBL) according to one embodiment of the present invention.

Detailed description of the invention

The present invention relates to a composition comprising at least one or more phytochemicals that can be used to effectively prevent and/or treat inflammation in humans and animals from natural sources that are thermally processed, such as extruded, such as plant material.

Applicants have surprisingly found that, by thermal processing, some plants and/or their plant extracts, such as chicory, can produce enzymes that are sensitive to cyclooxygenase enzymatic activity and/or NF-κB enhanced transcription in mammals Active phytochemicals with enhanced inhibitory effects, etc., which are believed to reduce the risk of inflammation, including, for example, dermatitis, ophthalmia, enteritis, colitis, and the like. In one embodiment, the enzymatic activity is from a cyclooxygenase, such as cyclooxygenase-1 and/or cyclooxygenase-2.

Applicants have demonstrated through tests that thermal processing, such as extrusion, of plant extracts, especially chicory extracts, inhibits cyclooxygenase activity as well as transcriptional activity of NF-κB. In particular, heat-processed (eg extruded) chicory extracts are believed to have a more pronounced effect on the inhibition of cyclooxygenase-2 than cyclooxygenase-1. Accordingly, inhibition of such enzymatic activity is believed to reduce inflammation in mammals. By reducing inflammation, the risk of developing other diseases or ailments believed to result from inflammation, especially chronic inflammation, is believed to be reduced. Other types of disease can include, for example, cancer, autoimmune disease, osteoarthritis, and combinations thereof.

In addition to the phytochemicals discussed above, plants such as chicory are also known to contain prebiotic fibers, such as oligosaccharides, including inulin, which are believed to reduce the incidence of cancer, especially in the colon. In this regard, applicants believe that enhanced benefits for cancer prevention and/or treatment in humans and animals can be achieved due to the combined effect of prebiotic fibers and phytochemicals that inhibit enzyme activity to prevent And/or treating inflammation in mammals, particularly chronic inflammation which can lead to cancer, such as colon cancer, if the inflammation is not addressed.

Applicants have also demonstrated that the enzyme inhibitory properties of thermally processed (eg extruded) compositions of the invention are not substantially affected by the processing conditions under which the compositions were prepared according to the invention. For example, the purification of plant material according to the invention, which can be used to reduce the bitterness of plant extracts and thus increase the palatability of humans and animals, has negligible impact, if any, on the nutritional properties of the resulting purified product . In this regard, the resulting extracted product may be derived from a substantially crude plant extract, such as chicory. This may preclude costly purification or other similar treatments of the plant material to produce the bioactive fraction of interest.

The term "biologically active agent" or other similar terms such as "bioactive fraction" as used herein means any component or components capable of exhibiting biological activity, chemical activity or similar activity in mammals, It enhances the health of mammals. Examples of bioactive agents include, eg, prebiotic fibers, phytochemicals, and the like.

The term "prebiotic" or other similar terms including "prebiotic fiber" as used herein means a substance or component that promotes the growth of microorganisms in a mammal.

As used herein, the term "phytochemical" or other similar terms including "phytochemical" and "phytochemical agent" means a plant-produced substance that is believed to confer health benefits on humans and/or animals such as the prevention and/or treatment of inflammation or any other chemical substance that resembles a disease.

As used herein, the term "enzymatic activity" or other similar terms such as "enzymatic activity" means any suitable enzyme that acts as a catalyst in any suitable biological, chemical, or other similar process that is thought to affect Mammalian health. For example, inhibition of enzymatic activity involving cyclooxygenase is thought to reduce the risk of inflammation.

As used herein, the term "thermal processing" or other similar terms such as "extrusion", "extruded", "extruded" means in a special equipment such as an oven or extruder, or can increase the The heating of plant raw materials and/or plant extracts above a standard temperature (such as 25 degrees Celsius or 278 Kelvin) in any similar device that processes the temperature of the material.

The compositions may include any suitable and compatible types and amounts of components such that the compositions are effective for preventing and/or treating inflammation. In one embodiment, the composition comprises plant material comprising one or more prebiotic fibers and a phytochemical that inhibits enzymatic activity, such as that involving cyclooxygenase. This is thought to treat and/or prevent inflammation. A number of plant materials can be effectively added to the composition including, for example, chicory, lettuce, coffee, soybean, Jerusalem artichoke, leek, asparagus, extracts thereof and combinations thereof. In one embodiment, chicory and/or chicory extracts are preferred.

It should be understood that the plant material can be processed in a number of different and suitable ways to form the extract. Typically, plant material, such as chicory root, is ground, pulverized or supplied in any suitable form. The plant material is then further processed through a number of different stages to produce the product extract. In one embodiment, a delipidation step is performed on the plant material to produce an extract resulting from the removal of fat from the plant material. The degreasing step may be performed using any suitable type and amount of solvent, including, for example, hexane, under any suitable degreasing process conditions.

In one embodiment, the extract obtained from the defatting step can be further processed by acid hydrolysis to produce another type of plant extract which can be added to the compositions of the present invention. The acid hydrolysis step can be performed using any suitable type and amount of solvent, including, for example, ethyl acetate, under any suitable process conditions.

In one embodiment, the extract obtained from the defatting step may be further processed by a solvent extraction step. Solvent extraction can be performed in the presence of any suitable amount and type of solvent, under any suitable process conditions. In one embodiment, the solvent comprises a solution of methanol ("MeOH") and water mixed 1:1 by volume. The solution obtained in the solvent extraction step can be further processed to produce another extract by evaporation of the solvent under suitable conditions. Alternatively, the resulting solution can be treated with an adsorbent such as polyvinylpolypyrrolidone or the like to capture polyphenols. Sorbent treatment can be carried out under any suitable process conditions. Specific examples of plant extracts prepared in accordance with embodiments of the present invention are described in detail below.

In one embodiment, the ground plant material is thermally processed. Applicants have demonstrated that, in this way, plant raw materials naturally comprising sesquiterpene lactones (SQLs) are enriched in the highly active COX2 inhibitor, α-methylene-γ-butyrolactone (α-MGBL). Indeed, this molecule exhibits enhanced thermal stability compared to other SQLs. For example, such molecules have been shown to inhibit inflammatory activity in part by directly interacting with the transcription factor NF-κB, inhibiting its binding to DNA. The inhibitory effect is proposed to result from the alkylation of amino acid Cys ³⁸ in the p65 domain of NF-κB. This inhibition results in decreased expression of several pro-inflammatory and inflammatory receptors and enzymes therein, including cyclooxygenase-II. Usually SQLs and α-MGBL specifically inhibit COX-2 activity and/or its expression. Applicants have demonstrated that α-CH ₂ -γ-butyrolactone specifically and highly inhibits COX-2 activity.

In one embodiment, the prebiotic fiber and phytochemical of the composition may be derived from common or the same plant material, such as chicory. As previously discussed, Applicants believe that the combined effects of prebiotic fiber and phytochemical sources of plants such as chicory may result in enhanced chemoprotection that allows inflammation to be treated and/or prevented in mammals. Prebiotic fibers may include any suitable amount and type, including, for example, oligosaccharides such as inulin and various fructan-oligosaccharides, soybean oligosaccharides, and combinations thereof.

Phytochemicals can include any suitable type and amount such that they inhibit COX-II enzymatic activity or NF-κB transcriptional activity, which is believed to prevent and/or treat inflammation. In one embodiment, the phytochemical of the plant material is capable of inhibiting cyclooxygenase activity such as cyclooxygenase-1 and/or cyclooxygenase-2 and/or NF-κB transcriptional activity. By inhibiting enzymatic and/or transcriptional activity, the phytochemicals of the invention are believed to prevent and/or treat inflammation, including, for example, chronic inflammation. In this regard, the risk of developing other diseases or ailments believed to be caused by inflammation such as cancer, osteoarthritis, autoimmune diseases may be reduced.

In addition to phytochemicals, plant sources may include other dietary ingredients that inhibit enzyme activity. In one embodiment, dietary ingredients include antioxidants, glucosamine, chondroitin sulfate, omega-3 fatty acids, the like, and combinations thereof.

It should be understood that the compositions of the invention may comprise a wide variety of different and suitable forms. In one embodiment, the composition may include nutritional supplements, human and/or animal food preparations, pet food and/or pharmaceutical and/or functional food compositions, and the like. The compositions can be added to food products in any suitable amount. In one embodiment, the food product comprises at least 0.5% by weight, preferably from about 1% to about 30% by weight, more preferably from about 1% to about 2% by weight, of the plant material composition.

In another embodiment, the pharmaceutical composition comprising the active product (such as α-methylene-γ butyrolactone, its product or extract) may be in any form suitable for oral use such as tablet , troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions for oral administration can be prepared according to any method used in the art for the production of pharmaceutical compositions, and such compositions can contain one or more selected from sweeteners, flavoring agents, coloring agents and preservatives to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with nontoxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets, such as inert diluents, granulating agents, disintegrants and lubricants. Tablets may be uncoated or coated using known techniques to delay disintegration and absorption in the gastrointestinal tract and thus provide sustained action over a longer period of time. Formulations for oral use may be hard gelatin capsules, in which the active ingredient is admixed with an inert solid diluent, or soft gelatin capsules, in which the active ingredient is admixed with an aqueous or oily vehicle. Aqueous suspensions contain the active materials in admixture with excipients such as suspending, dispersing or wetting agents, preservatives, coloring, flavoring and sweetening agents suitable for the manufacture of aqueous suspensions. Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Other excipients, for example sweetening, flavoring and coloring agents, may also be present.

In one embodiment, the present invention provides a pet food product comprising a starch base and an effective amount of plant material, wherein the plant material comprises prebiotic fiber and a phytochemical that inhibits an enzyme or enzymatic activity thereby enhancing mammalian Health, such as preventing and/or treating inflammation. The pet food products of the present invention may include any suitable number, type and amount of components and may be processed in any suitable manner to form the desired product form.

In one embodiment, the present invention includes a gelled cereal product comprising an amount of plant material. The plant material includes at least a source of prebiotic fiber and a source of phytochemicals capable of inhibiting enzyme activity in a mammal to enhance the health of the mammal.

In one embodiment, the plant includes inulin sufficient to provide at least about 0.25% inulin by weight, on a dry matter basis. The plant material used may be of any suitable source, for example chicory, lettuce, coffee, soybeans, Jerusalem artichokes, leeks, onions, yacon, asparagus and mixtures of these plants containing high levels of inulin. In one embodiment, chicory and Jerusalem artichoke are preferred. In one embodiment, the plant material comprises at least 50% by weight inulin. For ease of processing, the plant material is preferably in dry and comminuted or powder form. As described below, the method utilizes dried, comminuted chicory and/or extracts thereof. However, it should be understood that any suitable plant material may be used in any suitable form and added to the cereal product in any suitable amount.

As described below, the other ingredients included in the gelled cereal product may be any suitable ingredient commonly used in gelled cereal products. Typically these ingredients include a starch source and a protein source. Suitable sources of starch are, for example, cereals such as corn, rice, wheat, sugar beets, barley, oats, soybeans and mixtures thereof. Suitable protein sources may be selected from any suitable animal or vegetable protein source. Examples include meat meal, bone meal, fish meal, soy protein concentrate, dairy protein, gluten, and the like. The choice of starch source and protein source depends primarily on the nutritional requirements of the animal or human, palatability factors, the type of grain product being produced, or other factors. Various other ingredients such as sugar, salt, spices, seasonings, vitamins, minerals, flavorings, fats, etc. may also be incorporated into the gelled cereal product as desired.

Gelatinized cereal products can be produced in many different ways as desired. However, for dry cereal products, a particularly suitable way of producing the product is extrusion cooking. This can be done by methods well known in the art. For example, in one suitable method, the feed mixture is charged to a preconditioner. The feed mix consists mainly of a starch source, a protein source and plant material such as chicory. In one embodiment, chicory comprises at least about 1% by weight of the feed material, preferably at least about 2% by weight. In one embodiment, the amount of plant material in the food material ranges from about 10% to about 20% by weight, preferably about 10% by weight.

In the preconditioner, water or steam or both are mixed into the feed mixture. A sufficient quantity of water or steam is mixed into the feed mixture to moisten the feed mixture. If desired, the temperature of the feed mixture in the preconditioner can be raised to about 60°C to about 90°C by weight. A suitable preconditioner is described in US 4,752,139. It should be understood that the use of a preconditioner is not required.

The moist feed leaving the preconditioner is then loaded into an extruder. The extruder may be any suitable single or twin screw retort extruder. Suitable extruders are available from Wenger Manufacturing Inc., Clextral SA, Buhler AG, etc. On its way through the extruder, the moist feed passes through the cooking zone. In the cooking zone, the feed is mechanically sheared and heated. In one embodiment, the moistened feed is heated to a maximum temperature of up to about 150°C and passed through the forming zone. The gauge pressure of the forming zone is from about 300 kPa to about 10 MPa as desired. Water or steam or both can be introduced into the cooking zone if desired. During passage through the extruder, the starch source of the moist feed is gelatinized to provide a gelled matrix structure of primarily starch, protein and plant material such as chicory.

The gelling matrix leaving the extruder is forced through a suitable die, such as that described in EP0665051. A shaped extrudate exits the die, the extrudate having a profile consistent with the die orifice. The shaped extrudate expands to a greater or lesser extent depending on the conditions in the extruder and the starch source used. The shaped extrudate is then cut with a blade. The individual pieces are then dried and, if desired, coated with a preservative or flavoring or both. After cooling, the flakes can be packed into suitable packaging. Alternatively, individual chips can be flaked and then dried.

Depending on the ingredients used, the gelled cereal product may be in the form of a dry kibble suitable as a pet food, a bulked piece suitable for use in a breakfast cereal, a flake suitable for use in a breakfast cereal, etc.

It is also possible to produce dry cereal products by mixing water and ingredients of the cereal product together, eg in a preconditioner. The wet mixture is then formed into the desired shape, such as by using forming rolls. The shaped mixture is then baked in an oven at any suitable temperature. In one embodiment, the temperature is in the range of about 220°C to about 280°C for a suitable time. In one embodiment, the baking time ranges from about 10 minutes to about 1 hour. The dry cereal product has the appearance of a baked biscuit.

If it is desired to produce a simulated meat product for use in canned pet food, any suitable method may be used. For example, methods may include those described in US 4,781,939 and 5,132,137. In these methods, a protein source, particularly meat material, is emulsified. The meat material may be any suitable animal protein source including, for example, mammalian muscular or skeletal meat, poultry and fish or meat by-products such as heart, liver, kidney, tongue, etc. or meat meal. Vegetable protein sources can also be included if desired. Strict compositions can be chosen based on cost and desired taste. Emulsification can be performed in any suitable equipment.

Dried chicory is added to the emulsion. Other proteins may also be added to the emulsion if desired or necessary. The other protein may be any of the protein sources mentioned above. The strict choice will depend on availability, cost and palatability. Other proteins may be added in any suitable amount. In one embodiment, other proteins may be added in an amount ranging from about 5% to about 35% by weight.

Fat may also be added to the emulsion if desired or necessary. Usually the amount of fat in the emulsion must be controlled for ease of processing and to obtain an acceptable product. However, the meat material well contains the required amount of fat and therefore adjustments may not be necessary. Typically, at this point the emulsion contains a maximum fat level of about 25% by weight. In one embodiment, the amount of fat in the emulsion ranges from about 5% to 15% by weight, more preferably from about 7% to about 12% by weight. The mass ratio of protein to fat is preferably from about 1:1 to about 7:1. If fat is added, the fat may be any suitable animal fat such as beef tallow, or may be vegetable fat.

Other ingredients such as sugar, salt, spices, seasonings, flavorings, minerals etc. may also be added to the emulsion. In one embodiment, the amount of other ingredients used ranges from about 1% to about 5% by weight of the gelatinous cereal product.

Water may also be added to provide from about 45% to 80% by weight humidity in the emulsion. If sufficient humidity exists in the meat material, it is not necessary to add water.

Once mixed, the emulsion is preferably loaded into a vacuum packer or similar degassing equipment to degas the emulsion. This removes air which would otherwise cause disintegration of the formulated emulsion product and weaken its meaty appearance. The emulsion is then loaded into an emulsifier, subjecting the emulsion to rapid mechanical heating and shearing. Any suitable emulsifying machine including, for example, that disclosed in U.S. 5,132,137 may be used. Other suitable emulsifiers are commercially available under the trade name TRIGONAL and from Siefer Machinenfabrik GmbH & Co KG. Bahnhofstrasse 114, Postfach 101008., Velbert 1, Germany.

The temperature of the emulsion in the emulsifier can be raised to the desired coagulation temperature within seconds. In one embodiment, the condensation temperature ranges from about 100°C to about 120°C. In one embodiment, the temperature ranges from about 45°C to about 75°C as described in U.S. 5,132,137. Usually, the mechanical energy generated in the emulsifier is sufficient to heat the emulsion to the required temperature, but this can be supplemented by injecting superheated steam.

The heated emulsion leaving the emulsifier can be transferred into a collection tube. In the collecting tube, the heated emulsion coagulates while moving slowly along the collecting tube. The heated emulsion is present in the collection tube long enough for the emulsion to coagulate into a hard emulsion product when it reaches the outlet of the collection tube. The firm emulsion product exiting the collection tube is then transferred into a cutting machine where it is cut into chunk sizes suitable for use in pet food, such as chunks. Chunks have the shape and texture of meat. Chunks can be compressed into sheets if desired. Chunks can also be made as a chunk-in-gravy product topped with a bouillon sauce. Other methods of producing chunks are known and can be used, such as extruding the feed mix, cooking the feed mix on a steam oven and cutting the cooked extrudate into chunks.

If it is desired to produce canned pet food in the form of meat chunks, the meat paste can be prepared by emulsifying a suitable meat material to produce a mince. The meat material may be any suitable meat source, eg as described above. Suitable gelling agents including gums such as kappa carrageenan, carob bean gum, guar gum, xanthan gum, etc. may be added to the meat paste. In one embodiment, no more than about 2% by weight gum is used. Dried plant material such as chicory is then added to the mince.

Other ingredients such as sugar, salt, spices, seasonings, flavorings, minerals, etc. may also be added to the minced meat. The other ingredients are used in amounts such that they constitute from about 0.25% to about 5% by weight of the roux. Water may also be added to the ground meat to provide from about 70% to about 85% by weight. If sufficient humidity exists in the meat material, it is not necessary to add water.

The ground meat is then heated to above about 65°C in the mixer-cooker. Inject steam into the roux if desired. The heated ground meat is then re-emulsified to provide a loaf batter and the temperature of the loaf batter is maintained above about 60° C. until filled into tanks.

It should be understood that gelatinized cereal products may be produced by any suitable method and not only those described above. Other types of oligosaccharides such as oligofructosaccharides and soy oligosaccharides may also be included in the gelled cereal product. Soy oligosaccharides can be added in the form of soy flour or other suitable soy sources.

The cereal product may be in any suitable form including, for example, dry, semi-moist and moist. However, the matrix making up the cereal product must be gelled to remove or destroy the sesquiterpenes that may be present in the plant material. It should be understood that the cereal products of the present invention may be intended for human and/or animal consumption.

By way of illustration and not limitation, an example of a pet food product made according to one embodiment of the present invention is set forth below.

Example 1: Dry Pet Food

The feed mixture consisted of about 58% by weight corn, about 6% by weight corn gluten, about 23% by weight meat meal, with dried chicory, salt, vitamins and minerals making up the remainder. Dried chicory is added in the form of a chicory extract made according to one embodiment of the present invention and in an amount of about 5% or less. The feed mixture is loaded into the preconditioner and moistened. The moist feed is then loaded into the extruder-digester and gelled. As the gelled matrix exits the extruder, it is forced through a die and compressed to form an extrudate. The extrudate was cut into dog-friendly pieces, dried at about 110°C for about 20 minutes, and cooled to form pellets. It should be appreciated that part or all of the fat mixture, or part or all of the fats and oils used may be added at a later stage, eg as a coating.

Added Chicory As previously discussed, can enhance the health of pets by, for example, preventing and/or treating inflammation.

Example 2: Dry Pet Food

Dry pet food was prepared as the dry pet food of Example 1. It also includes other ingredients that are generally associated with enhancing the palatability of dry pet foods suitable for cats. As previously discussed, added chicory can enhance your pet's health by, for example, preventing and/or treating inflammation.

Example 3: Dry Cat Food

The feed mixture consisted of about 58% by weight corn, about 6% by weight corn gluten, about 23% by weight chicken meal, with dried chicory, salt, vitamins and minerals making up the remainder. Add about 5% or less chicory. As previously discussed, added chicory inhibits enzymatic activity, which is believed to enhance the health of the animal by, for example, treating and/or preventing inflammation.

The feed mixture is loaded into the preconditioner and moistened. The moist feed is then loaded into the extruder-digester and gelled. As the gelled matrix exits the extruder, it is forced through a die and compressed to form an extrudate. The extrudate was cut into cat-friendly pieces, dried at about 110°C for about 20 minutes, and cooled to form pellets. At this time, Lactobacillus and Enterococcus faecium such as Lactobacillus rhamnosus NCC2583 (CNCM I-2449), Lactobacillus acidophilus NCC2628 (CNCM I-2453) Freeze-dried powder of one or more strains of SF68 (NCIMB 10415) was applied to pellets. A sufficient amount of powder is applied to the pellets so that the corresponding dietary intake for cats is about 1.0E+07 to about 1.0E+9 colony forming units/day. At this point, a portion of the powder is mixed into the first batch of pellets which are then bagged. A second portion of powder is weighed and mixed with a liquid carrier, and then sprayed onto a second batch of pellets. The pellets were bagged after the coating had fully dried at 50-60°C for several minutes.

Example 4: Canned Pet Food and Supplements

The mixture is prepared from about 73% of livestock meat, pig lung and beef liver (ground), about 16% of wheat flour, about 2% of dyes, vitamins and inorganic salts. The mixture was emulsified at 12°C and extruded into pudding form. Dried chicory in the form of an extract made according to one embodiment of the present invention is added to the emulsification in an amount of about 5% or less. The emulsion was then cooked at 90°C. It is cooled to 30°C and cut into thick pieces. About 45% of the chunks are mixed with about 55% of the seasoning made from about 98% of water, about 1% of dye, and about 1% of guar gum. Fill tins with chunks and seasoning mixture and sterilize at 125°C for about 40 minutes.

Prebiotic supplements to be mixed with pet food before being served as a serving are provided in extra packs in the form of sachets with the following strain of Lactobacillus: Lactobacillus rhamnosus NCC2583 (CNCM I-2449 ), Lactobacillus acidophilus NCC2628 (CNCM1-2453) or Enterococcus faecium SF68 (NCIMB 10415). The corresponding dietary intake of supplements for pets is approximately 10 ⁶ -10 ¹² colony forming units/day, depending on the type of pet such as cat or dog, and on physical factors such as body weight of the pet. The supplement is packaged so that it is removably attached to the can, and there are feeding instructions.

By way of example and not limitation, the experimental tests used to demonstrate the efficacy of the present invention are described in detail below.

In vitro test 1

The inventors have carried out many experimental tests to demonstrate the beneficial effects of the present invention. Typically, human colon cancer cell cultures were prepared and treated with different amounts of chicory extract for about 48 hours to evaluate the effect of chicory on cyclooxygenase activity. Preparations and experimental testing procedures and results are discussed in more detail below.

Preparation of cell culture

Colon cancer cell line HT-29 was obtained from the American Type Culture Collection and cultured in McCoy's 5A medium supplemented with 10% fetal bovine serum (FBS), 25 mg/ml gentamicin. HT-29 cells were treated with various chicory extracts made according to one embodiment of the present invention as detailed below.

Preparation of Chicory Extract

Four different chicory extracts, Extracts A-D, were prepared according to one embodiment of the present invention. Initially, 40 grams (g) and 10 grams of powdered chicory samples were sieved through a 0.5 mm sieve, respectively. The fat was then removed by mixing the samples with hexane for about 30 minutes at room temperature. 600 milliliters (ml) of hexane was added to a 40-gram sieved sample of chicory, and 150 milliliters of hexane was added to a 10-gram sieved sample. The hexane was evaporated under vacuum at about 50°C to form extract A.

Extract B was prepared by first defatting 40 grams of ground chicory powder as discussed previously. The degreased sample is hydrolyzed with 300 ml of an acid, such as hydrochloric acid, for about 20 minutes in a boiling water bath. After cooling and centrifugation (8000 rpm, 5 minutes, 10° C.), the solution is extracted with 150 ml of a solvent such as ethyl acetate, which is commercially available, eg from MERCK. The solvent was evaporated to dryness. Extract B was formed after further drying over anhydrous sodium sulfate at about 50°C under vacuum.

Extracts C and D were prepared as follows. First, 10 grams of ground chicory powder was defatted as previously discussed. This second portion of skimmed powder is extracted with about 250 ml of a solvent/water mixture, such as a 1:1 volume ratio of MeOH and water. The extraction is carried out with stirring at room temperature (eg, about 20°C to about 25°C) for about 30 minutes. After centrifugation, the solution was divided into two equal volumes. For the first volume of the solution, the organic solvent was evaporated in vacuo at about 50°C. The remaining aqueous phase was lyophilized to form extract C.

A second volume of the solution was treated with about 2 grams of polyvinylpolypyrrolidone with stirring for about 30 minutes to capture polyphenols. The adsorbent material is removed by filtration, centrifugation, and the like. The organic solvent was evaporated in vacuo at about 50°C. The remaining aqueous phase was lyophilized to form extract D.

Determination of PGE2 production

The effect of various chicory extracts, Extracts A-C, on the biosynthesis of PGE2 was analyzed in the human colonic cell line HT-29. The biosynthesis of PGE2 provides a characterization of the level of enzymatic activity, such as cyclooxygenase, which is known to act as a catalyst in the production of PGE2 from, for example, arachidonic acid. Thus, the effect of chicory extract on cyclooxygenase enzyme activity can be evaluated based on the experiments described below.

Cells were grown for 48 hours in medium supplemented with 0.1% bovine serum albumin (BSA) and 10 [mu]M arachidonic acid. Chicory extract was then added to these media at concentrations of 50, 100 and 200 μg/ml for 21 hours or 15 hours. Another set of cell samples was prepared by adding chicory extract B to HT-29 cells as discussed above, followed by co-incubation in the presence of 10 ng/ml tumor necrosis factor alpha (TNF) for 6 hours. It is generally believed that the addition of TNF will stimulate inflammation.

The amount of PGE2 in the cell culture medium was determined using the PGE2 Monoclonal Enzyme Immunoassay Kit (Cayman Chemical) according to the manufacturer's instructions. Briefly, 25 or 50 microliters of medium and serially diluted PGE2 standards were mixed with appropriate amounts of acetylcholinesterase-labeled tracer and PGE2 antiserum and incubated at room temperature for 18 hours. After the wells were emptied and washed with wash buffer, 200 microliters of Ellman's reagent containing the substrate for acetylcholinesterase was added. Enzyme reactions were performed for 1 hour at room temperature on a slow shaker. Results were measured at 415 nm using a microplate reader and normalized to micrograms of protein.

The test results showed that chicory extract B prepared with ethyl acetate, as discussed above, had the most pronounced effect on the inhibition of cyclooxygenase activity, as evidenced by a reduction in the amount of PGE2 measured. In general, the decrease in cyclooxygenase activity was more pronounced with increasing amounts of chicory extract. In addition, the test results showed that chicory extract B had a more significant effect on the inhibition of cyclooxygenase activity in HT-29 cell line cultured with TNF than on the inhibition of cyclooxygenase activity in HT-29 cell line without TNF. This suggests that inhibition of cyclooxygenase activity is more strongly influenced by specific inhibition of cyclooxygenase-2 than cyclooxygenase-1. The results are listed in Table 1.

Table 1. Effect of Chicory Extract with Ethyl Acetate on PGE-2 Synthesis in HT-29 Cells Additives PGE-2 level (% control) - TNFα PGE-2 level (% control) + TNF-α ^* Control (vehicle only) 100 100 Chicory Extract B (100mcg/ml) 95 80 Chicory Extract B (100mcg/ml) 33 0.7

^* enzyme activity stimulated by TNF-α is 10 times

In vitro test 2

1. Inhibitory effect of α-CH ₂ -γ-butyrolactone on PGE2 synthesis

The present inventors have carried out similar experimental tests to demonstrate the anti-inflammatory effect of α-methylene-γ-butyrolactone. In general, the inhibitory effect of α-methylene-γ-butyrolactone on PGE2 synthesis has been shown in HT29 cells stimulated by TNF-α. For this, the cells were cultured and passaged in McCoy's 5A medium supplemented with arachidonic acid (10 microliters) and bovine serum albumin (0.01%). Cells were treated with α-methylene-γ-butyrolactone (in methanol) at doses of 15, 30, 60 μM or vehicle alone for 21 h with or without TNF-α (10 ng/ml ) stimulation for 6 hours. PGE2 analysis was performed using an Elisa test on media samples.

Table 2: In HT29 cells treated with α-CH ₂ -γ-butyrolactone with or without TNF-α

Concentration of PGE2 in pg/ml culture medium pgPGE ₂ /ml medium-TNF-α % control pgPGE ₂ /ml medium+TNF-α % control Methanol control α-CH ₂ -γ-butyrolactone 15 μM α-CH ₂ -γ-butyrolactone 30 μM α-CH ₂ -γ-butyrolactone 60 μM 2594281419701144 1001087614 189411328693491310 73051236050

2. Inhibitory effect of α-CH ₂ -γ-butyrolactone on the expression of COX-2

Western blot analysis: HT29 cells (treated as above for PGE- 2 assay, ie in the presence or absence of TNF-[alpha] and with or without [alpha] _-CH2-[ gamma]-butyrolactone) Cell lysates were prepared. Cell lysates were sonicated for 5 seconds and centrifuged at 10000 g for 5 minutes. Protein concentrations were determined by the Bradford method calibrated with bovine serum albumin.

Whole cell extracts (25 micrograms) were boiled in electrophoresis (Laemmli) sample buffer and separated by SDS-polyacrylamide gel electrophoresis on a 10% gel using a discontinuous polyacrylamide gel system. Non-specific binding sites were blocked by overnight incubation of membranes at 4°C in 5% dry milk phosphate buffer containing 0.1% tween-20.

Goat polyclonal anti-COX-2 and COX-1 antibodies (C-20, Santa Cruz Biotechnology) diluted 1:1000 in PBS containing 5% milk were incubated for 1 hour at room temperature for blot hybridization, and then blotted with 1% milk Wash with PBS for 1 hour.

Subsequently, the blot was probed with a donkey anti-goat secondary antibody conjugated to horseradish peroxidase. After washing for 1 hr, complexes were detected with ECL Western blotting reagents (Amersham). The results are shown in Figure 1.

3. Specific inhibitory effect of α-CH ₂ -γ-butyrolactone on COX-2 activity

Stressgen StressXpress™ Cyclooxygenase Activity Kit provides a method for the specific detection of cyclooxygenases COX-1 and COX-2 in various biological fluids. The kit uses a specific chemiluminescence substrate (aromatic hydrocarbon molecule) to detect the peroxidation activity of COX enzyme. After inhibition with specific compounds (α-CH2-γ-butyrolactone and NS-938, specific COX-2 inhibitors from Cayman Chemical Company, both dissolved in methanol), by adding chemiluminescent substrates and arachidene Acid (50 μM) was used to detect the immediate residual activity of cyclooxygenase. Light emission was started immediately and after 30 min the chemiluminescent signal (relative luminescence units - RLU) was detected (photometer Tecan). The amount of light emission is directly proportional to COX activity.

In this test, α-CH2-γ-butyrolactone was found to inhibit COX-2 activity as specifically and efficiently as the reference compound NS-398. The results are listed in Table 3.

Table 3: Determination of Cyclooxygenase (COX-1, COX-2) Activity COX-1 activity (RLU) % control COX-2 activity (RLU) % control Carrier only (methanol) 4115 100 26450 100 α-CH2-γ-butyrolactone (40μM) 4807 117 12823 49 NS-398(1mM) 4063 99 1794 7

It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and changes can be made without departing from the spirit and scope of the invention and without diminishing its intended advantages. It is therefore intended to cover such changes and changes in the appended claims.

Claims (33)

1. A composition comprising a therapeutically effective amount of plant material, wherein said plant material is thermally processed and includes one or more phytochemicals that inhibit at least one enzymatic activity and/or transcriptional activity To prevent inflammation in mammals. 2. A composition comprising a therapeutically effective amount of thermally processed plant material, wherein said plant material comprises one or more phytochemicals capable of inhibiting at least one enzymatic activity and/or transcriptional activity to treat inflammation in a mammal, Wherein the phytochemical agent includes an effective amount of sesquiterpene lactone, and the sesquiterpene lactone includes an active product containing α-methylene-γ-butyrolactone. 3. A composition comprising an active product derived from thermally processed plant material, the active product comprising α-methylene-γ-butyrolactone, wherein an effective amount of the active product can inhibit at least one enzyme and transcriptional activity to prevent or reduce inflammation. 4. A composition according to any one of claims 1 to 3, wherein plant material comprises at least 0.5% by weight. 5. A composition according to any one of claims 1 to 3, wherein the plant material comprises an effective amount of a sesquiterpene lactone including α-methylene-γ-butyrolactone-containing active product. 6. A composition according to any one of claims 1 to 3, wherein the plant material is from the family Asteraceae. 7. Composition according to any one of claims 1 to 3, wherein the plant material is derived from plants selected from the group consisting of coffee, soybean, chicory, lettuce, their extracts, their pulp and combinations thereof. 8. A composition according to any one of claims 1 to 3, wherein the plant material comprises chicory extract. 9. The composition according to any one of claims 1 to 3, wherein the plant material further comprises a dietary ingredient selected from the group consisting of antioxidants, glucosamine, chondroitin sulfate, omega-3 fatty acids, and combinations thereof. 10. The composition according to any one of claims 1 to 3, wherein the phytochemical is capable of inhibiting at least one of the enzymatic activity from cyclooxygenase and the transcriptional activity from NF-κB. 11. Composition according to any one of claims 1 to 3, wherein the composition is selected from nutritional supplements, nutritionally complete food products, food preparations, cereal products, pet food and/or pharmaceutical and/or or a functional food composition. 12. A composition according to any one of claims 1 to 3, wherein the thermally processed plant material comprises extruded plant material. 13. A pet food product comprising: a starch base and a therapeutically effective amount of thermally processed plant material comprising a phytochemical capable of inhibiting at least one enzyme and transcriptional activity in a mammal to reduce the risk of inflammation . 14. The pet food product of claim 13, wherein the thermally processed plant material comprises an effective amount of a sesquiterpene lactone comprising an active product comprising alpha-methylene-gamma-butyrolactone . 15. The pet food of claim 13, wherein the plant material is a plant material according to claims 4 to 9. 16. A pet food product comprising an effective amount of a plant material comprising an effective amount of a sesquiterpene lactone including α-methylene-γ-butyrolactone from a thermally processed plant material The active product, the plant material is selected from plants related to Compositae chicory, lettuce, coffee, soybean, their extracts, their pulp and their combination, to prevent or reduce inflammation. 17. A method of producing a nutraceutical food product capable of reducing the risk of inflammation in a mammal, comprising the steps of: providing plant material; processing the plant material to form a plant extract comprising one or more phytochemicals, said phytochemical an agent capable of inhibiting at least one enzyme activity and transcriptional activity in a mammal; and, processing the plant extract and one or more food ingredients to form a nutraceutical food product comprising at least 0.5% by weight of the plant extract. 18. The method of claim 17, wherein the plant material is thermally processed to provide an effective amount of a sesquiterpene lactone comprising an active product comprising alpha-methylene-gamma-butyrolactone . 19. The method of claim 17, wherein the plant material is extruded. 20. The method of claim 17, wherein the plant material is derived from a plant extract selected from the group consisting of coffee, soybean, chicory, lettuce, extracts thereof, pulps thereof, and combinations thereof. 21. The method of claim 17, wherein the plant extract is processed by defatting the plant material to form the primary plant extract, and the primary plant extract is subsequently processed by extraction with ethyl acetate and acid hydrolysis to form the plant extract . 22. The method of claim 17, wherein the plant extract further comprises a dietary ingredient selected from the group consisting of antioxidants, glucosamine, omega-3 fatty acids, and combinations thereof. 23. Use of a heat-processed therapeutically effective amount of a composition comprising plant material comprising a phytochemical capable of inhibiting at least one enzymatic and transcriptional activity in a mammal, prepared for use as a treatment for an effective Composition for reducing the risk of inflammation in mammals at risk of inflammation. 24. Use of a therapeutically effective amount of a composition comprising thermally processed plant material comprising a phytochemical capable of inhibiting at least one of an enzymatic activity and a transcriptional activity in a mammal, prepared for use as a treatment for an effective A composition for reducing the risk of osteoarthritis in a mammal at risk of osteoarthritis. 25. Use of a therapeutically effective amount of a composition comprising thermally processed plant material comprising a phytochemical capable of inhibiting at least one of an enzymatic activity and a transcriptional activity in a mammal, prepared for use as a treatment for an effective A composition for reducing the risk of an autoimmune disease in a mammal at risk of developing an autoimmune disease. 26. Use of a therapeutically effective amount of a composition comprising thermally processed plant material comprising a phytochemical capable of inhibiting at least one enzymatic and transcriptional activity in a mammal, prepared for use as a treatment for a patient with A composition for reducing the risk of cancer in a mammal at risk of cancer. 27. Use according to any one of claims 23 to 26, wherein the plant material is from a plant selected from the group consisting of Compositae coffee, soybean, chicory, lettuce, their extracts, their pulp and combinations thereof. 28. Use according to any one of claims 23 to 26, wherein the phytochemical is capable of inhibiting at least one of an enzymatic activity involving cyclooxygenase and a transcriptional activity involving NF-[kappa]B. 29. Use according to any one of claims 23 to 26, wherein the plant material comprises a plant extract from chicory. 30. The use according to any one of claims 23 to 26, wherein the thermally processed plant material comprises an effective amount of a sesquiterpene lactone comprising α-methylene-γ-butanol Active product of lactones. 31. Use according to any one of claims 23 to 26, wherein the composition comprises at least 0.5% by weight of plant material which has been thermally processed and contains an effective amount of a sesquiterpene lactone, which Esters include reactive products containing α-methylene-γ-butyrolactone. 32. Use of a therapeutically effective amount of [alpha]-methylene-[gamma]-butyrolactone, or components or extracts comprising it, for the preparation of a nutritional or pharmaceutical composition for inhibiting COX-2 activity. 33. Use according to claim 32, wherein the nutritional or pharmaceutical composition reduces the risk of inflammation, osteoarthritis or autoimmune disease or cancer in the mammal.